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  1. Link

    erk1-antibody-ep4967-ab109282.pdf

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Neuroscience Neurology process Neurodegenerative disease Alzheimer's disease Tangles & Tau
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ERK1 antibody [EP4967] (ab109282)

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Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] (ab109282)
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [EP4967] (ab109282)
  • Flow Cytometry (Intracellular) - Anti-ERK1 antibody [EP4967] (ab109282)
  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] (ab109282)
  • Anti-ERK1 antibody [EP4967] (ab109282)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP4967] to ERK1
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-ERK1 antibody [EP4967]
    See all ERK1 primary antibodies
  • Description

    Rabbit monoclonal [EP4967] to ERK1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293T, Jurkat, A375, A431 and HUVEC cell lysates. IHC-P: Human colon adenocarcinoma tissue. Flow Cyt (intra): Jurkat cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP4967
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Tangles & Tau
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic

Associated products

  • Alternative Versions

    • Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human MAPK3 (ERK1) knockout HEK-293T cell line (ab266519)
  • KO cell lysates

    • Human MAPK3 (ERK1) knockout HEK-293T cell lysate (ab257099)
  • KO cell pellets

    • Human MAPK3 (ERK1) knockout HEK-293T cell pellet (ab278963)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)
    • A-431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant human ERK1 protein (ab105904)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109282 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/30.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000 - 1/10000. Predicted molecular weight: 44 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.

ICC/IF
1/100.
Notes
Flow Cyt (Intra)
1/30.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000 - 1/10000. Predicted molecular weight: 44 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Heat up to 98°C, below boiling, and then let cool for 10-20 min.

ICC/IF
1/100.

Target

  • Function

    Involved in both the initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors such as ELK-1. Phosphorylates EIF4EBP1; required for initiation of translation. Phosphorylates microtubule-associated protein 2 (MAP2). Phosphorylates SPZ1 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4).
  • Sequence similarities

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • Domain

    The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • Post-translational
    modifications

    Dually phosphorylated on Thr-202 and Tyr-204, which activates the enzyme. Dephosphorylated by PTPRJ at Tyr-204.
  • Target information above from: UniProt accession P27361 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5595 Human
    • Entrez Gene: 26417 Mouse
    • Entrez Gene: 50689 Rat
    • Omim: 601795 Human
    • SwissProt: P27361 Human
    • SwissProt: Q63844 Mouse
    • SwissProt: P21708 Rat
    • Unigene: 861 Human
    • Unigene: 8385 Mouse
    • Unigene: 2592 Rat
    see all
  • Alternative names

    • ERK 1 antibody
    • ERK-1 antibody
    • ERK1 antibody
    • ERT 2 antibody
    • ERT2 antibody
    • Extracellular Signal Regulated Kinase 1 antibody
    • Extracellular signal related kinase 1 antibody
    • Extracellular signal-regulated kinase 1 antibody
    • HGNC6877 antibody
    • HS44KDAP antibody
    • HUMKER1A antibody
    • Insulin Stimulated MAP2 Kinase antibody
    • Insulin-stimulated MAP2 kinase antibody
    • MAP kinase 1 antibody
    • MAP kinase 3 antibody
    • MAP Kinase antibody
    • MAP kinase isoform p44 antibody
    • MAPK 1 antibody
    • MAPK 3 antibody
    • MAPK antibody
    • MAPK1 antibody
    • Mapk3 antibody
    • MGC20180 antibody
    • Microtubule Associated Protein 2 Kinase antibody
    • Microtubule-associated protein 2 kinase antibody
    • Mitogen Activated Protein Kinase 3 antibody
    • Mitogen-activated protein kinase 1 antibody
    • Mitogen-activated protein kinase 3 antibody
    • MK03_HUMAN antibody
    • OTTHUMP00000174538 antibody
    • OTTHUMP00000174541 antibody
    • p44 ERK1 antibody
    • p44 MAPK antibody
    • p44-ERK1 antibody
    • p44-MAPK antibody
    • P44ERK1 antibody
    • P44MAPK antibody
    • PRKM 3 antibody
    • PRKM3 antibody
    • Protein Kinase Mitogen Activated 3 antibody
    see all

Images

  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : MAPK3 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 44 kDa
    Observed band size: 43 kDa why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab109282 observed at 43 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab109282 was shown to react with ERK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266519 (knockout cell lysate ab257099) was used. Wild-type HEK-293T and MAPK3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109282 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] (ab109282)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] (ab109282)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling ERK1 with purified ab109282 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [EP4967] (ab109282)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [EP4967] (ab109282)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat (human acute T cell leukemia) cells labelling ERK1 with purified ab109282 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Flow Cytometry (Intracellular) - Anti-ERK1 antibody [EP4967] (ab109282)
    Flow Cytometry (Intracellular) - Anti-ERK1 antibody [EP4967] (ab109282)

    Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling ERK1 with purified ab109282 at 1/30 dilution (red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control. 

     

     

  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    Western blot - Anti-ERK1 antibody [EP4967] (ab109282)

    Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Green signal from target - ab109282 observed at 42 kDa
    Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
    Lanes 5 and 6: Merged (red and green) signal
    ab109282 was shown to specifically react with ERK1 when ERK1 knockout samples were used.
    Wild-type and ERK1 knockout samples were subjected to SDS-PAGE. ab109282 and ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    Western blot - Anti-ERK1 antibody [EP4967] (ab109282)

    Predicted band size : 43 kDa

    Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Green signal from target 
    Lanes 3 and 4: Red signal from loading control 
    Lanes 5 and 6: Merged (red and green) signal

    Red - loading control, ab8226 observed at 42 kDa or ab8245, observed at 37 kDa

    This western blot image is a comparison between ab109282 and a competitor's top cited rabbit polyclonal antibody.

  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/10000 dilution (purified)

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate
    Lane 3 : NIH/3T3 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 44 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/10000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : A375 cell lysate
    Lane 3 : HUVEC cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 44 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    Western blot - Anti-ERK1 antibody [EP4967] (ab109282)
    All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/1000 dilution (unpurified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : A375 cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : HUVEC cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 44 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] (ab109282)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] (ab109282)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colonic adenocarcinoma tissue labelling ERK1 with unpurified ab109282 at 1/100 dilution.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Anti-ERK1 antibody [EP4967] (ab109282)
    Anti-ERK1 antibody [EP4967] (ab109282)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab109282? Please let us know so that we can cite the reference in this datasheet.

ab109282 has been referenced in 2 publications.

  • Zhi Q  et al. Podocalyxin-like protein promotes gastric cancer progression through interacting with RUN and FYVE domain containing 1 protein. Cancer Sci 110:118-134 (2019). PubMed: 30407695
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371

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